Efficient Parthenogenesis Induction and In Vitro Haploid Plant Regeneration in Cucumber (Cucumis sativus L.) Using Putrescine, Spermidine, and Cycocel
- 219 Downloads
In this study, the effect of spraying mother plants with various levels of putrescine, spermidine, and cycocel (each at 0, 50, 500, and 5000 mg/l) were assessed on the frequency of haploid embryos produced from unfertilized ovaries and subsequent regeneration of derived embryos. Significantly higher haploid embryos were obtained when mother plants were sprayed with putrescine at 500 mg/l (5.2 embryos/fruit), spermidine at 50 mg/l (4.8 embryos/fruit), and cycocel at 50 mg/l (5.2 embryos/fruit) as compared to the control (without spraying, 3.2 embryos/fruit). However, embryogenesis induction was decreased drastically as the concentration of all the three compounds tested was increased and the lowest haploid embryos were observed when 5000 mg/l of spermidine (0.4 embryos/fruit) or cycocel (2.0 embryos/fruit) were applied. Only spermidine at 50 mg/l led to 100% regeneration into fully developed plantlets. The seed setting and size of fruits were also affected by polyamines and cycocel applications. Ploidy analysis using a flow cytometer indicated that all regenerated plantlets contain the gametic chromosome number (n = x = 7) of parental plants and the results of chromosome counting also confirmed the haploid nature of regenerated plantlets. It can be concluded that the induction of haploid embryogenesis from unfertilized ovaries after pollination with irradiated pollen and subsequent conversion of derived embryos into the plantlets could be improved in Cucumis sativus L. by applying appropriate levels of putrescine, spermidine, and cycocel.
KeywordsCucumis sativus L. Embryo rescue Haploid Parthenogenesis Polyamines
This research was supported by grants from Agricultural Biotechnology Research Institute of Iran (ABRII) Project No. 12-05-05-9451-94001.
Compliance with Ethical Standards
Conflict of interest
The authors declare that they have no conflict of interests.
- Alifar M, Ebadi A, FatahiMoghadam MR (2015) Effects of putrescine and chlormequat pre-flowering treatment on embryo rescue technique success in diallele crosses of grape cultivars Flame Seedless, perlette and Yaghooti. Iran J Hortic Sci 46(2):179–192Google Scholar
- Altinas S (2011) Effects of chlormequat chloride and different rates of prohexadione-calcium on seedling growth, flowering, fruit development and yield of tomato. Afr J Biotechnol 10(75):17160–17169Google Scholar
- Ebrahimi R, Zamani Z (2009) Effect of polyamines on in vitro gynogenesis of onion (Allium cepa L.). Am Eur J Sustain Agric 3(1):71–74Google Scholar
- El Meskaoul A, Trembaly FM (2009) Effects of exogenous polyamines and inhibitors of polyamine biosynthesis on endogenous free polyamine contents and the maturation of white spruce somatic embryos. Afr J Biotechnol 8(24):6807–6816Google Scholar
- Gałązka J, Niemirowicz-Szczytt K (2013) Review of research on haploid production in cucumber and other cucurbits. Folia Hortic 25(1):67–78Google Scholar
- Kamiab F, Salehabad MH, Zamani bahramabadi E (2015) Evaluation the effects of foliar treatments of polyamines and some organic acids on quantitative and qualitative traits in some pistachio cultivars. J Nuts 6(2):131–142Google Scholar
- Kumar HGA, Ravishankar BV, Murthy HN (2004) The influence of polyamines on androgenesis of Cucumis sativus L. Eur J Hortic Sci 69(5):201–205Google Scholar
- Lewitsky GA (1931) An essay on cytological analysis of the fixing action of the chrom-acetic formalin and the chromic formalin. Bull Appl Bot 27(1):181–185Google Scholar
- Miroshnichenko D, Filippov M, Doglov S (2009) Effects of daminozide on somatic embryogenesis from immature and mature embryos of wheat. Aust J Crop Sci 3(2):83–94Google Scholar
- Mukherjee A, Bandyopadhyay A (2014) Inducing somatic embryogenesis by polyamines in medicinally important Clerodendrum indicum L. Int J Curr Microbiol App Sci 3(8):12–26Google Scholar
- Ponce MT, Guinazo M, Tizio R (2002) Effect of putrescine on embryo development in the stenospermocarpic grape cvs Emperatriz and Fantasy. Vitis 41(1):53–54Google Scholar
- Saleem BA, Malik AU, Anwar R, Farooq M (2006) Exogenous application of polyamines improves fruit set, yield and quality of sweet oranges. Acta Hortic 774:187–194Google Scholar
- Shariatpanahi ME, Ahmadi B (2016) Isolated microspore culture and its applications in plant breeding and genetics. In: Anis M, Ahmad N (eds), Plant tissue culture: propagation, conservation and crop improvement. Springer Science + Business Media Singapore, pp 487–507Google Scholar
- Thiruvengadam M, Rekha KT, Jayabalan N, Praveen N, Kin EH, Chung IM (2013) Effect of exogenous polyamines enhances somatic embryogenesis via suspension cultures of spine gourd (Momordica dioica Roxb. ex. Willd.). Aust J Crop Sci 7(3):446–453Google Scholar
- Wei A, Du S, Han Y, Zhang G (2010) A study on the relationship between cucumber gynogenesis and content of ovary hormones and polyamines. Acta Hortic 871:625–630Google Scholar